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Confocal Raman Microscopy for Label-Free Detection of ProteinLigand Binding at Nanopore-Supported Phospholipid Bilayers David A. Bryce, Jay P. Kitt, and Joel M. Harris Anal. Chem., Just Accepted Manuscript • DOI: 10.1021/acs.analchem.8b02791 • Publication Date (Web): 03 Sep 2018 Downloaded from http://pubs.acs.org on September 3, 2018
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Analytical Chemistry
Confocal Raman Microscopy for Label-Free Detection of Protein-Ligand Binding at Nanopore-Supported Phospholipid Bilayers David A. Bryce, Jay P. Kitt, and Joel M. Harris* Department of Chemistry, University of Utah, 315 South 1400 East, Salt Lake City, UT 84112-0850 USA ABSTRACT Interactions of lectins, proteins that selectively bind carbohydrates, play an important role in many biological processes including cell-adhesion, immune response, and cell signaling. Given the range of lectin functions and their potential for application in disease-detection, there is a need for methods to investigate lectin-carbohydrate interactions that are rapid, structurally-specific, and sensitive to binding from low-concentration samples. In this work, we describe preparation and application of supported phospholipid bilayers deposited in wide-pore chromatographic silica particles for confocal Raman microscopy-based detection of specific binding of concanavalin-A to mannose-functionalized phospholipids. The high surface area of porous-silica supports provides an ample concentration of phospholipid and protein for rapid, label-free detection of lectin binding to be carried out in an individual lipid-bilayerfunctionalized particle. The Raman spectrum provides structural information on the bound protein as well as the phospholipid bilayer. Using scattering from the supported-lipid bilayer as an internal-standard, Raman scattering from accumulated protein can be interpreted quantitatively to determine its absolute surface coverage on the lipid bilayer. At low glycolipid fraction (
